芪仙通络方及其拆方对脑梗死大鼠室管膜下区神经发生及p38MAPK活化的影响  

Effects of Qixian Tongluo Prescription and Its Disassembled Prescriptions on Neurogenesis and p38MAPK Activation in Subependymal Zone of Rats with Cerebral Infarction

作　　者：周胜强 李博[3] 王琦 周春吉 刘芳 邓奕辉[2] ZHOU Shengqiang;LI Bo;WANG Qi;ZHOU Chunji;LIU Fang;DENG Yihui(Hunan Academy of Chinese Medicine,Changsha 410006,China;Hunan University of Chinese Medicine,Changsha 410208,China;The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;The First Affiliated Hospital of University of South China,Hengyang 421001,China)

机构地区：[1]湖南省中医药研究院,湖南长沙410006 [2]湖南中医药大学,湖南长沙410208 [3]湖南中医药大学第一附属医院,湖南长沙410007 [4]南华大学第一附属医院,湖南衡阳421001

出　　处：《中国中医药信息杂志》2021年第9期85-91,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家中医药管理局“刘祖贻国医大师传承工作室”建设项目(2015年);湖南省教育厅重点项目(18A225);湖南省教育厅优秀青年项目(18B253);湖南省中医药研究院院级重点课题(201902);湖南省“国内一流培育学科”中西医结合开放基金项目(2019ZXYJH01)。

摘　　要：目的观察芪仙通络方及其拆方对脑梗死大鼠室管膜下区神经发生及p38MAPK活化的影响,探讨其作用机制。方法SD大鼠随机分为假手术组、模型组、芪仙通络方组(全方组)、补肾活血拆方组(拆方1组)、益气温阳拆方组(拆方2组)和胞磷胆碱组,线栓法制备大鼠大脑中动脉阻塞模型,造模后第3日开始灌胃给药,同时腹腔注射5-溴脱氧尿嘧啶核苷(BrdU)标记脑内增殖细胞,每日1次,连续12 d。于造模后第3、7、14日对大鼠神经功能进行评分;免疫荧光染色观察大鼠缺血侧室管膜下区巢蛋白(Nestin)/BrdU、双肾上腺皮质激素(doublecortin,DCX)/BrdU、神经元核抗原(NeuN)/BrdU双标阳性细胞数;免疫组化检测大鼠缺血侧室管膜下区磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)蛋白表达;Pearson相关分析法分析Nestin/BrdU、DCX/BrdU和NeuN/BrdU双标阳性细胞数与神经功能评分和p-p38MAPK蛋白表达的相关性。结果与假手术组比较,模型组大鼠各时间点神经功能评分明显升高(P<0.01),大鼠缺血侧室管膜下区Nestin/BrdU、DCX/BrdU和NeuN/BrdU双标阳性细胞数明显增加(P<0.05),p-p38MAPK蛋白表达升高,但差异无统计学意义(P>0.05);与模型组比较,各给药组大鼠第7、14日神经功能评分明显降低(P<0.01),大鼠缺血侧室管膜下区Nestin/BrdU、DCX/BrdU、NeuN/BrdU双标阳性细胞数明显增多(P<0.01),p-p38MAPK蛋白表达显著上调(P<0.01),全方组作用最显著(P<0.05)。全方组大鼠缺血侧室管膜下区Nestin/BrdU、DCX/BrdU、NeuN/BrdU双标阳性细胞数与神经功能评分呈显著负相关,与p-p38MAPK蛋白表达呈显著正相关。结论芪仙通络方及其拆方可改善脑梗死大鼠受损神经功能,其机制可能与促进缺血侧室管膜下区p38MAPK活化、上调内源性神经发生水平有关。Objective To observe the effects of Qixian Tongluo Prescription and its disassembled prescriptions on neurogenesis and p38MAPK activation in the subependymal zone in rats with cerebral infarction;To explore its mechanism.Methods The SD rats were randomly divided into sham-operation group,model group,Qixian Tongluo Prescription group(full prescription group),Bushen Huoxue disassembled prescription group(disassembled prescription group 1),Yiqi Wenyang disassembled prescription group(disassembled prescription group 2),and Citicoline group.The suture method was used to replicate the middle cerebral artery occlusion model in rats.The intragastric administration was started on the 3rd day after modeling,and 5-bromodeoxyuridine(BrdU)was injected intraperitoneally to label proliferating cells in the brain once a day for 12 consecutive days.On the 3rd,7th,and 14th day after modeling,neurological function was scored;immunofluorescence staining was used to observe ischemic side subependymal zone Nestin/BrdU,doublecortin(DCX)/BrdU,neuronal nuclei antigen(NeuN)/BrdU double-labeled positive cells;immunohistochemistry was used to detect the expression of phosphorylated p38 mitogen-activated protein kinase(p-p38MAPK)in the subependymal zone of the ischemic side of rats;Pearson correlation analysis method was used to analyze the correlation between the number of Nestin/BrdU,DCX/BrdU and Nestin/BrdU double-labeled positive cells with neural function score and p-p38MAPK protein expression.Results Compared with the sham-operation group,the neurological function scores of the model group increased significantly at each time point(P<0.01);the number of Nestin/BrdU,DCX/BrdU and NeuN/BrdU double-labeled positive cells in the subependymal zone of the ischemic side of the rat increased significantly(P<0.05);the expression of p-p38MAPK protein increased,without statistical significance(P>0.05).Compared with the model group,the nerve function scores of rats in each administration group were significantly reduced on the 7th and 14th day(P<0.

关 键 词：芪仙通络方 脑梗死 神经发生 P38丝裂原活化蛋白激酶 大鼠 

分 类 号：R285.5[医药卫生—中药学]